Polypeptides play an important role in today's medical portfolio. For human application every pharmaceutical substance has to meet distinct criteria. To ensure the safety of biopharmaceutical agents to humans nucleic acids, viruses, and host cell proteins, which would cause severe harm, have to be removed especially. To meet the regulatory specification one or more purification steps have to follow the manufacturing process. Among other purity, throughput, and yield play an important role in determining an appropriate purification process.
In general chromatographic methods chromatography columns are employed which are essentially comprising a column housing with an upper and lower fitting which in turn comprises an inlet at the top of the column, an outlet at the bottom of the column, a upper chromatography column frit, a lower chromatography column frit, an upper distributor plate and a chromatographic material.
General chromatographic methods and their use are known to a person skilled in the art. See for example, Chromatography, 5th edition, Part A: Fundamentals and Techniques, Heftmann, E. (eds), Elsevier Science Publishing Company, New York, (1992); Advanced Chromatographic and Electromigration Methods in Biosciences, Deyl, Z. (ed.), Elsevier Science BV, Amsterdam, The Netherlands, (1998); Chromatography Today, Poole, C. F., and Poole, S. K., Elsevier Science Publishing Company, New York, (1991); Scopes, Protein Purification: Principles and Practice (1982); Sambrook, J., et al. (eds), Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989; or Current Protocols in Molecular Biology, Ausubel, F. M., et al. (eds), John Wiley & Sons, Inc., New York.
For the purification of immunoglobulins, which have been produced e.g. by cell cultivation methods, in general a combination of different chromatography steps is employed. Normally a protein A affinity chromatography is followed by one or two additional separation steps. The final purification step is a so called “polishing step” for the removal of trace impurities and contaminants like aggregated immunoglobulins, residual HCP (host cell protein), DNA (host cell nucleic acid), viruses, or endotoxins. For this polishing step normally an anion exchange material in a flow-through mode is used.
In WO 2006/048514 a container comprising open, complementarily-shaped upper and lower ends with a chromatographic mixture which is placed in the container, and a membrane filter which is fixed to the lower end of the container is reported. A high throughput liquid chromatography column assembly including a loading column with a loading chamber with a first inner diameter and a first length in fluid communication with a separation column having a separation chamber with a diameter smaller than the loading column's inner diameter and a length greater than the loading column's length is reported in U.S. Pat. No. 6,458,273.
High pressure liquid chromatography columns are reported in U.S. Pat. No. 4,719,011 which may be modularly modified as to length and/or internal diameter and may contain other components in the modular system, for example, column sections, adapters and cone adapters, for joining column sections of different internal diameters, and end plate units for funneling in or discharging out fluids. A column device comprising a stationary phase having a plurality of particles adapted for interacting with a mobile phase in order to separate different compounds of a sample fluid dissolved in the mobile phase, a housing for at least partly housing the stationary phase, and a separator separating sections of the stationary phase and being force-coupled with the housing is reported in EP 1 916 522.
A segmented chromatography column including a plurality of media beds separated and bounded by a plurality of porous members is reported in WO 00/010675. In U.S. Pat. No. 3,398,512 a chromatography apparatus is reported. Chromatography columns with partition elements therein are reported in U.S. Pat. No. 3,453,811. In EP-A 1 892 526 a column and cartridge column using the same is reported. A separation system for the resolving of volatile mixtures is reported in U.S. Pat. No. 3,657,864.